U.S. Pat. No. 4,402,950 is considered as the closest prior art as it discloses (in vitro) the antiviral activity of carvone against adenovirus type 6, which is a double stranded DNA non enveloped virus.
U.S. Pat. No. 4,402,950 discloses in its claim 1 “a process for deactivating viruses inside living human and animal organisms infected with said viruses comprising administering to one of said organisms a terpene selected from the group consisting of black pepper oil, cinnamon flower oil, cardamon oil, linallyl acetate, cinnamic aldehyde, carvone, and cis/trans citral, in a dosage amount effective to deactivate said viruses but ineffective to cause toxic effects on living cells of the living organism.”
The first column of U.S. Pat. No. 4,402,950 mentions at line that the carvone used is from the fruit of Carum carvi. The man skilled in the art will therefore easily deduct that the carvon used in U.S. Pat. No. 4,402,950 is the enantiomer “(+)-carvon”. In fact it is well known in the literature that S-(+)-Carvone is the principal constituent of the oil from caraway seeds (Carum carvi).
The difference between the present invention and the closest prior art is:
a composition comprising R-(−)-2-methyl-5-(prop-1-en-2-yl)-cyclohex-2-enone (also called (−)-carvon) and (2E)-3,7-dimethylocta-2,6-dien-1-ol (also called trans-geraniol) in combination with at least one more component chosen among essential oils in a pharmaceutically effective concentration (see list D which is disclosed in the present invention) for use in treatment and prevention of diseases caused by DNA enveloped viruses, DNA non-enveloped viruses, RNA enveloped viruses and RNA non-enveloped viruses, said diseases are those mentioned in claim 1.
The technical effect of the above mentioned difference is that an unexpected and surprising regression of lesions on the surface of the cervix occurred in vivo (between 20%-100%: see the comparative tests), while there is no medical effect occurring when any individual component is tested alone in vivo.
The objective technical problem to be solved is considered as the provision of a synergetic (alternative) antiviral composition comprising (+) carvone having an antiviral effect.
The question to be answered now is whether there is any teaching in the prior art as a whole (like U.S. Pat. No. 3,429,971) that would have prompted the skilled person, faced with the objective technical problem, to modify the closest prior art while taking account of that teaching, thereby arriving at something falling within the terms of the claims, and thus achieving what the invention achieves. The answer to this question is clearly NO. We explain why as follows:
U.S. Pat. No. 3,429,971 discloses a method of treating avian lymphomatosis induced by ES strain of lymphomatosis virus which comprises administering to a bird an effective amount of Cis-Geraniol. It is well known from a man skilled in the art that the isomers Cis-Geraniol and Trans-Geraniol have different biological/biochemical properties. This is even proved in U.S. Pat. No. 3,429,971:
The skilled person would never have combined U.S. Pat. No. 4,402,950 with U.S. Pat. No. 3,429,971 because U.S. Pat. No. 3,429,971 discloses in column 3, table 2 second and third rows:
“Trans-2,6-dimethyl-2,6,octadiene-8-ol→ No. cures: 0”.
Trans-2,6-dimethyl-2,6,octadiene-8-ol corresponds to the component “Trans-geraniol, i.e. to (2E)-3,7-dimethylocta-2,6-dien-1-ol”.
The virus mentioned in U.S. Pat. No. 3,429,971 is the avian lymphomatosis virus, which is a double stranded DNA enveloped virus (herpesviridae family).
The man skilled in the art, starting from U.S. Pat. No. 4,402,950 would be led to search a second component deactivating a virus of the same family as U.S. Pat. No. 4,402,950, i.e. a DNA non enveloped virus and therefore would not be led to search a further component deactivating a DNA enveloped virus. Even, if the man skilled in the art would be aware of the existence of U.S. Pat. No. 3,429,971, he would never use the component Trans-Geraniol in combination with carvone because column 3, table 2 second and third rows of U.S. Pat. No. 3,429,971 would dissuade him to combine both components because the man skilled in the art will learn that Trans-Geraniol does not have any curing effect (see table 2 of U.S. Pat. No. 3,429,971, namely the value zero cure).
In addition, the man skilled in the art would never find any indication in the prior art giving him any hint to use a composition containing carvone and Trans-Geraniol combined with at least one more component chosen among essential oils in a pharmaceutically effective concentration and selected from the list of components of claim 2 because the components listed in claim 2 are nor disclosed in U.S. Pat. No. 4,402,950, nor in U.S. Pat. No. 3,429,971, in combination.
The comparative tests listed in the present invention prove the above assertion. Even if the man skilled in the art would have, for an unknown reason, combined carvone with Trans-Geraniol for deactivating a DNA non enveloped virus, the comparative tests prove that no regression of lesions on the surface of the cervix have been observed, while for the synergetic composition of claim 1, 20%-100% of regression of lesions on the surface of the cervix have been observed. Consequently, there is a synergism between the combination of (+) carvone and (−) carvone and Trans-Geraniol and at least one more component chosen among essential oils in a pharmaceutically effective concentration of the present invention provoking an unexpected and surprising effect, i.e. 20%-100% of regression of lesions on the surface of the cervix.
Clinical and experimental pharmacology and physiology (2005) 32, pages 811-816 entitled “antiviral activities of extracts and selected pure constituents of ocimum basilicum” shows that purified components of ocimum basilicum were used to identify possible antiviral activities against human DNA viruses (HSV, ADV) and RNA viruses (CVB1, EV71). No activity was noted for carvone, cineole, beta-caryophylene, farnesol, fenchone, geraniol, beta-myrcene and alpha-thujone (it is to understand that no activity was noted for each component used alone). The difference between the present invention and this document is that the composition of the present invention is a synergetic combination of several compounds which proved having an unexpected antiviral effect.
Antiviral research 42 (1999) pages 219-226 is entitled “plant products as topical microbicide candidates: assessment of in vitro and in vivo activity against herpes simplex virus type 2” relates to protective efficacy in vivo in a mouse (see page 220 second column last paragraph) and a guinea pig model (see page 223 second column) of genital HSV-2 infection. At page 223 second column it is mentioned that infection in a guinea pig model, unlike in the mouse, does not result in death and more closely mimics the natural course of disease in humans. For these experiments, 0.1 ml of eugenol was instilled into the vaginal vault of female guinea pigs followed 20 seconds later by instillation of 0.1 ml of virus suspension containing 106 pfu of HSV-2 MS strain. Table 2 shows that treatment with eugenol immediately prior to intravaginal inoculation resulted in significant fewer animals developing symptomatic primary disease compared to PBS-treated controls. These data indicate that eugenol does have direct antiviral activity. The difference between the present invention and “Antiviral research 42 (1999) pages 219-226” is that in the present invention the novel synergetic composition is instilled after the viruses infected the human body in order to treat human diseases. It is obvious for a man skilled in the art that eugenol can be used as a prophylactic in vivo if it is instilled in a dosis of 0.1 ml prior to instillation of 0.1 ml of HSV-2 MS strain suspension in the vagina. The operating steps in the present invention are inversed comparing to those mentioned in the prior art document “Antiviral research 42 (1999) pages 219-226”. In addition, Eugenol in the present invention has the chemical formula “2-methoxy-4-prop-2-enylphenol” while page 223 first column second paragraph mentions eugenol as having the formula “4-hydroxy-3-methoxy-allylbenzene” which might be a different structural formula than that of the present invention.
Phytotherapy research 14, pages 495-500 (2000) entitled “In vitro and in vivo activity of Eugenol on human herpesvirus” mentions that Eugenol-acyclovir combinations synergistically inhibited herpesvirus replication in vitro and that topical application of eugenol delayed the development of herpesvirus induced keratitis in a mouse. Eugenol in the present invention has the chemical formula “2-methoxy-4-prop-2-enylphenol”, while in the abstract of this document eugenol is mentioned as having the formula “4-hydroxy-3-methoxy-allylbenzene” which might be a different structural formula than that of the present invention. The difference between the present invention and this document is that the composition of the present invention is a synergetic combination of several different compounds (other than the combination Eugenol-acyclovir) which proved having an unexpected antiviral effect. In addition, page 497 second column last paragraph bridging with page 498 first column first paragraph discloses that different dilutions of eugenol were instilled in each eye of mice before virus inoculation; after infection mice received the corresponding treatment three times a day and during 5 days (i.e. 15 applications). The difference between the present invention and this document is that in the present invention the novel synergetic composition is instilled after the viruses infected the human body in order to treat human diseases. The operating steps in the present invention are inversed comparing to those mentioned in this prior art document. It is obvious for a man skilled in the art that eugenol can be used as a prophylactic in vivo if it is instilled in a dosis of 1 mg/ml or 0.5 mg/ml prior to virus inoculation.
Natural Product Communications, 2008 vol. 3, No 7, 1085-1088 entitled “investigation of anticancer and antiviral properties of selected aroma samples” discloses that synthetic nerolidol samples at concentrations less than 5 microMolar have anti-tumor effects in vitro. Antiviral studies were performed to investigate inhibitory effects of the compounds on mouse polyomavirus (MPyV) propagation in 3T6 cells. 3T6 cells were washed with DMEM and incubated with virus inoculum for 1 h at 37° C. After that, DMEM supplemented with FBS and containing compounds to be analyzed was added. The difference between the present invention and this document is that the composition of the present invention is a synergetic combination of several different compounds which proved having an unexpected antiviral effect on humans. Results of tests on human beings cannot be compared to in vitro tests.
General Comments on the Prior Art Documents:
Any prior art document dealing with in vitro tests cannot be considered pertinent because the compositions of the present invention have been tested in vivo i.e. on animals or on human beings and it is well known by a man skilled in the art that test results can strongly differ if they are performed in an in vitro versus an in vivo environment. An extensive literature search did not find out any evidence or correlation that even if a composition is active in vitro, a man skilled in the art could deduct that the same composition would also be active in vivo. We also refer to Antiviral research 42 (1999) pages 219-226 is entitled “plant products as topical microbicide candidates: assessment of in vitro and in vivo activity against herpes simplex virus type 2” (already previously mentioned) where it is stated at page 224 first column “Several compounds showed activity in vitro but performed poorly in vivo. This failure of in vitro results to predict in vivo efficacy has previously been noted by zeitlin et al. (1997)”. This is even proven in the table of this document.